NLRP3在小鼠脓毒症相关性脑病中的作用及其与小胶质细胞焦亡的关系  

Role of NLRP3 in sepsis-associated encephalopathy and the relationship with pyroptosis in microglia of mice

作　　者：李翠[1] 董树安[1] 宋凯 李香云 杜诗涵 余剑波[1] Li Cui;Dong Shu′an;Song Kai;Li Xiangyun;Du Shihan;Yu Jianbo(Deportment of Anesthesiology and Critical Care Medicine,Tianjin Medical University NanKai Hospital(Tianjin Nankai Hospital),Tianjin 300100,China)

机构地区：[1]天津医科大学南开临床学院(天津市南开医院)麻醉科和重症医学科,天津300100

出　　处：《中华麻醉学杂志》2023年第2期206-209,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(82104584)。

摘　　要：目的评价核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)在小鼠脓毒症相关性脑病中的作用及其与小胶质细胞焦亡的关系。方法SPF级健康雄性C57BL/6J小鼠24只,6~8周龄,体质量18~22 g,采用随机数字表法分为3组(n=6):假手术组(Sham组)、脓毒症相关性脑病组(SAE组)和脓毒症相关性脑病+NLRP3抑制剂MCC950组(SAE+MCC950组)。采用盲肠结扎穿孔法制备小鼠脓毒症相关性脑病模型。SAE+MCC950组小鼠造模后1 h时腹腔注射MCC95020 mg/kg,余组注射等容量生理盐水。造模后1 d时行旷场实验,记录站立次数和中央区停留时间;造模后2~3 d行新物体识别实验,记录探索指数。造模后3 d行为学实验结束后,处死小鼠取脑海马组织,采用Western blot法检测NLRP3的表达,采用免疫荧光技术计数NLRP3与小胶质细胞特异性离子钙结合衔接分子1(Iba-1)共表达细胞,采用ELISA法检测caspase-1活性、IL-1β和IL-18含量。结果与Sham组比较,SAE组和SAE+MCC950组站立次数减少,中央区停留时间缩短,探索指数降低,海马NLRP3表达上调,NLRP3^(+)-Iba-1^(+)细胞计数增加,caspase-1活性、IL-1β和IL-18含量升高(P<0.05);与SAE组比较,SAE+MCC950组站立次数增多,中央区停留时间延长,探索指数升高,NLRP3表达下调,NLRP3^(+)-Iba-1^(+)细胞计数减少,caspase-1活性、IL-1β和IL-18含量降低(P<0.05)。结论NLRP3参与了小鼠脓毒症相关性脑病的发生,与其介导小胶质细胞焦亡有关。Objective To evaluate the role of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)in sepsis-associated encephalopathy(SAE)and the relationship with pyroptosis in microglia of mice.Methods Twenty-four SPF healthy male C57BL/6J mice,aged 6-8 weeks,weighing 18-22 g,were divided into 3 groups(n=6 each)using a random number table method:sham operation group(Sham group),SAE group and SAE plus an NLRP3 inhibitor MCC950 group(SAE+MCC950 group).The mouse model of SAE was prepared by cecal ligation and puncture after anesthesia.MCC95020 mg/kg was intraperitoneally injected at 1 h after developing the model in SAE+MCC950 group,and the equal volume of normal saline was given instead in the other groups.Open field tests were conducted at 1 day after developing the model to record the number of rearing and time spent in the central area.Novel object recognition tests were conducted at 2-3 days after developing the model to record the recognition index.After the behavioral experiment on 3 day after developing the model,mice were sacrificed and hippocampal tissues were collected for determination of the expression of NLRP3(by Western blot),count of cells co-expressing NLRP3 and microglia-specific ionized calcium-binding adaptor molecule 1(Iba-1)(by immunofluorescence),activity of caspase-1,and contents of interleukin-1beta(IL-1β)and IL-18(by enzyme-linked immunosorbent assay).Results Compared with Sham group,the number of rearing was significantly reduced,the time spent in the central area was shortened,the recognition index was decreased,the expression of NLRP3 was up-regulated,the count of NLRP3^(+)-Iba-1^(+)cells was increased,and the activity of caspase-1 and contents of IL-1βand IL-18 were increased in SAE and SAE+MCC950 groups(P<0.05).Compared with SAE group,the number of rearing was significantly increased,the time spent in the central area was prolonged,the recognition index was increased,the expression of NLRP3 was down-regulated,the count of NLRP3^(+)-Iba-1^(+)cells was decreased,and the activ

关 键 词：NLR家族 热蛋白结构域包含蛋白3 脓毒症相关性脑病 小神经胶质细胞 细胞焦亡 

分 类 号：R459.7[医药卫生—急诊医学]